Similar controls on calcification under ocean acidification across unrelated coral reef taxa

Ocean acidification (OA) is a major threat to marine ecosystems, particularly coral reefs which are heavily reliant on calcareous species. OA decreases seawater pH and calcium carbonate saturation state (Ω), and increases the concentration of dissolved inorganic carbon (DIC). Intense scientific effort has attempted to determine the mechanisms via which ocean acidification (OA) influences calcification, led by early hypotheses that calcium carbonate saturation state (Ω) is the main driver. We grew corals and coralline algae for 8–21 weeks, under treatments where the seawater parameters Ω, pH, and DIC were manipulated to examine their differential effects on calcification rates and calcifying fluid chemistry (Ω_cf, pH_cf, and DIC_cf). Here, using long duration experiments, we provide geochemical evidence that differing physiological controls on carbonate chemistry at the site of calcification, rather than seawater Ω, are the main determinants of calcification. We found that changes in seawater pH and DIC rather than Ω had the greatest effects on calcification and calcifying fluid chemistry, though the effects of seawater carbonate chemistry were limited. Our results demonstrate the capacity of organisms from taxa with vastly different calcification mechanisms to regulate their internal chemistry under extreme chemical conditions. These findings provide an explanation for the resistance of some species to OA, while also demonstrating how changes in seawater DIC and pH under OA influence calcification of key coral reef taxa.     

Response to technical comment on `meta-analysis reveals negative yet variable effects of ocean acidification on marine organisms'

It has been proposed that crustaceans should be excluded from a comparison of biological responses to ocean acidification among organisms with different calcium carbonate (CaCO3 ) forms in their calcified structures. We re-analysed our data without crustaceans and found high variation in organismal responses within CaCO3 categories. We conclude that the CaCO3 polymorph alone does not predict sensitivity, and a consideration of functional differences among organisms is necessary for predicting variation in response to acidification.     

Calcein labelling and electrophysiology: insights on coral tissue permeability and calcification

The mechanisms behind the transfer of molecules from the surrounding sea water to the site of coral calcification are not well understood, but are critical for understanding how coral reefs are formed. We conducted experiments with the fluorescent dye calcein, which binds to calcium and is incorporated into growing calcium carbonate crystals, to determine the permeability properties of coral cells and tissues to this molecule, and to determine how it is incorporated into the coral skeleton. We also compared rates of calcein incorporation with rates of calcification measured by the alkalinity anomaly technique. Finally, by an electrophysiological approach, we investigated the electrical resistance of coral tissues in order to better understand the role of tissues in ionic permeability. Our results show that (i) calcein passes through coral tissues by a paracellular pathway, (ii) intercellular junctions control and restrict the diffusion of molecules, (iii) intercellular junctions should have pores of a size higher than 13 Å and lower than 20 nm, and (iv) the resistance of the tissues owing to paracellular junctions has a value of 477 ± 21 Ohm cm². We discuss the implication of our results for the transport of calcium involved in the calcification process.     

PHACOTUS LENTICULARIS (CHLAMYDOMONADALES,PHACOTACEAE) ZOOSPORES REQUIRE EXTERNAL SUPERSATURATION OF CALCIUM CARBONATE FOR CALCIFICATION IN CULTURE1,2

In nature, zoospores of the chlamydophycean genus Phacotus Perty usually have a calcified lorica. The only cultured species, Phacotus lenticularis (Ehrenberg) Stein, did not readily calcify in artificial media. To overcome this deficiency, we developed an artificial culture medium (N-HS) in which Phacotus lenticularis formed mineralized loricae, as under natural conditions. Calcification of Phacotus (strain Krienitz 91/1) was achieved in a medium containing the ionic concentrations found in natural habitats (i.e. Lake Haussee and Lake Stechlin), hard-water lakes of the Baltic Lake District (Germany). The N-HS medium contained extremely low phosphate concentrations and high calcium and magnesium concentrations compared with common culture media, but the concentrations were similar to those in the lakes. Calcium carbonate in N-HS medium was dissolved up to the saturation concentration (saturation index = 1). Supersaturation was achieved by the addition of ultra-alkaline compounds (Na₂S_iO₃ or NaOH). The medium with the highest super-saturation had a saturation index of 118 and was extremely supersaturated with respect to calcium carbonate. In that medium and in modified media with less of the sodium compounds (lower supersaturations), calcification of zoospores was observed. To determine the effects of sodium and silicon compounds on calcification, various other silicon and sodium compounds were tested but were shown to be ineffective. In conclusion, calcification of Phacotus lenticularis depends directly on the degree of calcium carbonate supersaturation of the medium, but the fundamental mineralization pattern does not. Our study shows that calcification in Phacotus lenticularis can be triggered and controlled by supersaturation of calcium carbonate.     

Molecular analysis of two genes, DD9A and B, which are expressed during the postmolt stage in the decapod crustacean Penaeus japonicus

In decapod crustaceans, deposition of calcium carbonate crystals (calcification) in the exoskeleton takes place during the postmolt phase of the molt cycle. In an attempt to identify proteins which regulate the calcification process, the differential display technique was used to identify genes which were specifically expressed in the integument during the postmolt stage in the penaeid prawn Penaeus japonicus. One of the genes thus identified, named DD9A, was expressed in the epithelial cells of the tail fan. DD9A encoded a putative precursor of a secreted protein of 113 amino acids which exhibited sequence similarities to a group of crustacean and insect cuticular proteins, suggesting that DD9A was a protein component of the exoskeleton. Another gene, DD9B, which was also transcribed specifically during the postmolt period was identified based on its sequence similarity to DD9A. Potential roles of the DD9A protein in the calcification of the exoskeleton will be discussed.     

施肥方式和园龄对洛川苹果园土壤钙素退化的影响

为明确黄土高原苹果产区施肥方式和园龄对土壤钙素含量和钙素贮量的影响,本研究以位于世界苹果优生区的陕西省洛川县苹果园为研究对象,分别研究不同施肥方式和不同园龄苹果园0～100 cm土层土壤碳酸钙、水溶性钙和交换性钙含量及其贮量的变化特征.结果表明: 洛川县苹果园土壤钙素递减式退化现象严重,长期大量单施化肥苹果园土壤钙素退化现象明显大于化肥与农家有机肥配施苹果园,单施化肥苹果园比化肥与农家有机肥配施苹果园0～100 cm土层土壤碳酸钙、水溶性钙和交换性钙平均含量分别减少38.8%、25.4%和5.6%,3种形态土壤钙素贮量依次减少36.4%、26.0%和4.3%;苹果园土壤钙素退化程度随园龄增加不断加剧,园龄>25年苹果园比园龄≤10年苹果园0～100 cm土层土壤碳酸钙、水溶性钙和交换性钙平均含量分别减少48.8%、69.4%和39.5%,3种形态土壤钙素贮量分别减少40.8%、64.1%和33.0%.长期大量单施化肥和长期种植苹果树均对土壤碳酸钙、水溶性钙、交换性钙有明显的耗竭作用,钙素递减式退化特征明显,化肥与农家有机肥配施能够有效减缓土壤钙素退化,对于园龄>25年的高龄苹果园应加强土壤钙素管理.施肥方式是苹果园土壤钙素递减的驱动因素,钙素递减呈现出明显的时(园龄)空(土层深度)效应.     

Cyanobacterial formation of intracellular Ca‐carbonates in undersaturated solutions

Cyanobacteria have long been thought to induce the formation of Ca‐carbonates as secondary by‐products of their metabolic activity, by shifting the chemical composition of their extracellular environment to conditions favoring mineral precipitation. Some cyanobacterial species forming Ca‐carbonates intracellularly were recently discovered. However, the environmental conditions under which this intracellular biomineralization process can occur and the impact of cyanobacterial species forming Ca‐carbonates intracellularly on extracellular carbonatogenesis are not known. Here, we show that these cyanobacteria can form Ca‐carbonates intracellularly while growing in extracellular solutions undersaturated with respect to all Ca‐carbonate phases, that is, conditions thermodynamically unfavorable to mineral precipitation. This shows that intracellular Ca‐carbonate biomineralization is an active process; that is, it costs energy provided by the cells. The cost of energy may be due to the active accumulation of Ca intracellularly. Moreover, unlike cyanobacterial strains that have been usually considered before by studies on Ca‐carbonate biomineralization, cyanobacteria forming intracellular carbonates may slow down or hamper extracellular carbonatogenesis, by decreasing the saturation index of their extracellular solution following the buffering of the concentration of extracellular calcium to low levels.     

Bacterial Calcium Carbonate Precipitation in Cave Environments: A Function of Calcium Homeostasis

To determine if microbial species play an active role in the development of calcium carbonate (CaCO ₃ ) deposits (speleothems) in cave environments, we isolated 51 culturable bacteria from a coralloid speleothem and tested their ability to dissolve and precipitate CaCO ₃ . The majority of these isolates could precipitate CaCO ₃ minerals; scanning electron microscopy and X-ray diffractrometry demonstrated that aragonite, calcite and vaterite were produced in this process. Due to the inability of dead cells to precipitate these minerals, this suggested that calcification requires metabolic activity. Given growth of these species on calcium acetate, but the toxicity of Ca ²⁺ ions to bacteria, we created a loss-of-function gene knock-out in the Ca ²⁺ ion efflux protein ChaA. The loss of this protein inhibited growth on media containing calcium, suggesting that the need to remove Ca ²⁺ ions from the cell may drive calcification. With no carbonate in the media used in the calcification studies, we used stable isotope probing with C ¹³ O ₂ to determine whether atmospheric CO ₂ could be the source of these ions. The resultant crystals were significantly enriched in this heavy isotope, suggesting that extracellular CO ₂ does indeed contribute to the mineral structure. The physiological adaptation of removing toxic Ca ²⁺ ions by calcification, while useful in numerous environments, would be particularly beneficial to bacteria in Ca ²⁺ -rich cave environments. Such activity may also create the initial crystal nucleation sites that contribute to the formation of secondary CaCO ₃ deposits within caves.     

Carotenoid pigments and trophic behaviour of deep-sea shrimps (Crustacea, decapoda, alvinocarididae) from a hydrothermal area of the mid-atlantic ridge

Pigments and trophic behaviour of three species of Alvinocarididae from a Mid-Atlantic hydrothermal site were analysed. Carotenoid pigments are responsible for the more or less marked colouration of these animals. The carotenoid content of whole animals and different tissues were evaluated. Rimicaris exoculata exhibits an increased carotenoid level at the juvenile stage, while Chorocaris chacei and Alvinocaris markensis contain only few traces of pigment. Free and esterified astaxanthin, reported for most pelagic crustaceans, are present in these deep-sea shrimps. The origin of carotenoids of crustaceans living in the aphotic zone is discussed.     

The skeletal organic matrix from Mediterranean coral Balanophyllia europaea influences calcium carbonate precipitation

Scleractinian coral skeletons are made mainly of calcium carbonate in the form of aragonite. The mineral deposition occurs in a biological confined environment, but it is still a theme of discussion to what extent the calcification occurs under biological or environmental control. Hence, the shape, size and organization of skeletal crystals from the cellular level through the colony architecture, were attributed to factors as diverse as mineral supersaturation levels and organic mediation of crystal growth. The skeleton contains an intra-skeletal organic matrix (OM) of which only the water soluble component was chemically and physically characterized. In this work that OM from the skeleton of the Balanophyllia europaea, a solitary scleractinian coral endemic to the Mediterranean Sea, is studied in vitro with the aim of understanding its role in the mineralization of calcium carbonate. Mineralization of calcium carbonate was conducted by overgrowth experiments on coral skeleton and in calcium chloride solutions containing different ratios of water soluble and/or insoluble OM and of magnesium ions. The precipitates were characterized by diffractometric, spectroscopic and microscopic techniques. The results showed that both soluble and insoluble OM components influence calcium carbonate precipitation and that the effect is enhanced by their co-presence. The role of magnesium ions is also affected by the presence of the OM components. Thus, in vitro, OM influences calcium carbonate crystal morphology, aggregation and polymorphism as a function of its composition and of the content of magnesium ions in the precipitation media. This research, although does not resolve the controversy between environmental or biological control on the deposition of calcium carbonate in corals, sheds a light on the role of OM, which appears mediated by the presence of magnesium ions.     

Significance of the N- and C-terminal regions of CAP-1, a cuticle calcification-associated peptide from the exoskeleton of the crayfish, for calcification

Calcification-associated peptide (CAP)-1 is considered to play an important role in calcification of the exoskeleton of the crayfish, Procambarus clarkii. In this study, in order to clarify the molecular mechanism of calcification, we constructed expression systems of recombinant molecules of CAP-1 and its related peptides in Escherichia coli, and verified the structure-activity relationship of these peptides. The inhibitory assay on calcium carbonate precipitation and the calcium-binding assay showed that the C-terminal acidic region was most important for both activities. The CD spectra of these peptides varied depending on calcium concentration and showed that the N-terminal region is associated with the peptide conformation. These results indicate that the C-terminal part of CAP-1 may concentrate calcium ions for nucleation and/or interact with calcium carbonate precipitate to control the growth and that the N-terminal part contribute to maintaining the peptide conformation.     

Concrétions Minérales Intracytoplasmiques chez les Ciliés

SUMMARY. Various species of ciliates are characterized by the formation and accumulation in the cytoplasm of mineral concretions which are refringent, isotropic or anisotropic. These cytoplasmic inclusions most often are composed of calcium carbonate; in several species, however, their nature remains partially or even totally undetermined. The isotropic calcium-containing concretions often exhibit a definite shape; the calcium carbonate in this case appears to be bound to an organic substrate. The physiological role of the calcic concretions is not known; their characteristic presence in a given species is not necessarily related to ecological conditions. In a few species the calcification is localized in definite structures: spicules, skeletal plates, or otoliths of organelles supposedly sensory in nature.     

The relationship between calcification and photosynthesis in the coccolithophorid Pleurochrysis carterae

Coccolithophorids are one of the dominant groups of marine phytoplankton. They are found in large numbers throughout the surface euphotic zone of the ocean, and are able to form large-scale blooms that persist for long periods of time. Coccolithophorid cells are covered by species-specific calcium carbonate crystals of various structures. In the process of calcification in coccolithophorids, Ca²⁺ is absorbed into cells from the culture medium, and a coccolith unit is formed inside the cell. Then, the coccolith unit extrudes to the cell surface where it is constructed into crystal layers. The formation of these crystals is regulated by cellular metabolism under different environmental conditions. The carbon biogeochemical cycle in the coccolithophorids involves both photosynthetic and calcification processes, which not only play an important role in population dynamics, but also in the global carbon cycle and climate change. However, one important question remains, namely, whether the relationship between photosynthesis and calcification is species-dependent. Previous studies have yielded controversial results, even in the same species. In this paper, we selected Pleurochrysis carterae, a coccolithophore species that frequently blooms in coastal areas, to study the relationship between calcification and photosynthesis. First, we studied population growth in a batch culture over several days. For batch cultures, P. carterae was inoculated into a 10 L bioreactor at an initial cell density of approximately 5 × 10⁴ cells mL^-1. The culture conditions were optimal for cell growth. Dissolved oxygen (DO) was detected during all the culture period, and the rate of photosynthetic oxygen evolution was calculated according the DO changes during the 12-h illumination period. Algal samples (10 mL) were collected during the population growth phases. The calcium carbonate content on the cell surface was determined each day by chemical titration. Next, we studied the relationship between photosynthesis and calcification at the cellular level by observing patterns of recalcification during a 12-h period. In this study, non-calcified cells were obtained by decalcifying calcified cells collected during the exponential growth period in MES-NaOH buffer solution (pH 5.5). The non-calcified cells were inoculated into culture media containing different concentrations of Ca²⁺ (0, 5, 20, 40, 50, or 100 mg L^-1). The rate of recalcification was determined by microscopic analyses in which the number of recalcified cells per 100 cells was counted at 0, 3, 6, 9, and 12 h of culture. Ca²⁺ absorbed into the cell was detected by measuring the fluorescence intensity of Fluo-3/AM labeled Ca²⁺. The rate of photosynthetic oxygen evolution in the non-calcified cell cultures was detected by measuring the changes in dissolved oxygen during the 12-h illumination period. The results showed that during the population growth period, the rate of photosynthetic oxygen evolution was inversely related to the calcium carbonate content per cell. When the amount of calcium carbonate on the cell surface increased, the relative photosynthetic ability (the rate of photosynthetic oxygen evolution) decreased, and vice versa. Both recalcification rates and photosynthetic oxygen evolution were affected by the extracellular calcium concentration. Non-calcified cells showed different recalcification abilities at different extracellular Ca²⁺ concentrations. The recalcification rate of non-calcified cells was positively correlated with the extracellular calcium concentration when [Ca²⁺] in the medium ranged from 0 to 100 mg L^-1. However, photosynthetic oxygen evolution was suppressed at higher cell calcification rates, especially when extracellular [Ca²⁺] was 50–100 mg L^-1. Our analyses of the population growth process and the cell recalcification process confirmed that photosynthesis is inversely related to calcification in P. carterae.     

CALCIUM BALANCE AND MOULTING IN THE CRUSTACEA

1. Crustaceans have a high content of calcium, which is chiefly located in the skeleton as calcium carbonate. Calcium is generally the most abundant cation in the body. 2. During intermoult, the exoskeleton is usually fully calcified and the animal is in calcium equilibrium with its environment. 3. In the premoult stages calcium is resorbed from the skeleton and may be lost to the environment or stored within the body. Typically, losses are high and storage is small in aquatic species, whilst most terrestrial forms store much larger amounts of calcium and losses are reduced. Loss of calcium in soluble form by aquatic species must be by outward transport across the gills. 4. Calcium is stored in a variety of different ways, usually with a common taxonomic theme. The main forms are as calcium phosphate granules in cells of the midgut gland (Brachyura), gastroliths (Astacidea and some Brachyura), the haemocoel (some Brachyura) the posterior midgut caeca (Amphipoda) and the ventral portion of the body generally in the Isopoda. 5. At ecdysis, the skeleton is shed and the calcium remaining in it is lost from the body. 6. Recalcification begins immediately, or shortly after, ecdysis using calcium mobilized from the stores. Simultaneously, or when the stores are exhausted, other sources of calcium are utilized. These are calcium in the water (aquatic species), the food (aquatic and terrestrial species) and the exuviae (chiefly terrestrial species). 7. Marine species store little calcium and must obtain the bulk of their requirement (ca. 95%) from the water. Fresh-water species also store little calcium but have, seemingly, adapted to the lower availability of calcium by increasing the affinity of the calcium-absorbing mechanism. The rates of uptake of calcium are consequently similar in marine and fresh-water species. 8. A high degree of storage is essential for terrestrial crustaceans as they do not have access to a large aquatic reservoir of calcium. These large reserves enable the animals to reach an advanced stage of calcification, allowing the resumption of foraging and feeding necessary for completion of calcification. 9. The control of calcium metabolism during the intermoult cycle is poorly under stood. β Ecdysone appears to control the resorption of calcium and the formation of calcium stores during premoult, but the mechanism of control of calcium metabolism during postmoult and intermoult is unknown. 10. The concentration of calcium in the haemolymph of most species is high, but a large proportion of this is in non-ionized form. In premoult, total calcium levels rise as a result of calcium resorption but little change occurs in the concentration of ionized calcium. Postmoult generally sees a fall in blood calcium, sometimes below the intermoult value.     

Characterization of the exoskeleton of the Antarctic king crab Paralomis birsteini

Ocean acidification is projected to inhibit the biogenic production of calcium carbonate skeletons in marine organisms. Antarctic waters represent a natural environment in which to examine the long‐term effects of carbonate undersaturation on calcification in marine predators. King crabs (Decapoda: Anomura: Lithodidae), which currently inhabit the undersaturated environment of the continental slope off Antarctica, are potential invasives on the Antarctic shelf as oceanic temperatures rise. Here, we describe the chemical, physical, and mechanical properties of the exoskeleton of the deep‐water Antarctic lithodid Paralomis birsteini and compare our measurements with two decapod species from shallow water at lower latitudes, Callinectes sapidus (Brachyura: Portunidae) and Cancer borealis (Brachyura: Cancridae). In Paralomis birsteini, crabs deposit proportionally more calcium carbonate in their predatory chelae than their protective carapaces, compared with the other two crab species. When exoskeleton thickness and microhardness were compared between the chelae and carapace, the magnitude of the difference between these body regions was significantly greater in P. birsteini than in the other species tested. Hence, there appeared to be a greater disparity in P. birsteini in overall investment in calcium carbonate structures among regions of the exoskeleton. The imperatives of prey consumption and predator avoidance may be influencing the deposition of calcium to different parts of the exoskeleton in lithodids living in an environment undersaturated with respect to calcium carbonate.     

Shell formation and calcium transport in the barnacle Chthamalus fragilis

The mantle epithelium of the barnacle Chthamalus fragilis (Darwin) exhibits several ultrastructural features which may serve to regulate the calcification process. At the basis-mural plate and intermural plate junctions where rapid shell growth occurs, cells are characterized by long apical cytoplasmic projections and large intercellular spaces. These features may increase the functional surface area of the epithelium and enable more rapid deposition of calcium. The cells underlying the general shell surfaces contain numerous electron-dense inclusion bodies and show frequent cellular disintegration near the growing shell interface. Release of the granular contents of these inclusion bodies has been observed in both disintegrating and non-disintegrating cells. X-ray microanalysis revealed significantly higher calcium levels in the inclusion bodies than in the surrounding cytoplasm. This suggests a calcium transport role for these inclusion bodies. Cellular debris produced as a result of the disintegration of the mantle cells near the shell may play some role in the formation of the organic matrix of the shell. The presence of large numbers of mitochondria and well-developed apical microvilli in the cells of the inner mantle epithelium suggest that these cells serve to transport calcium into the mantle from the ambient sea water.     

Orchestin, a calcium-binding phosphoprotein, is a matrix component of two successive transitory calcified biomineralizations cyclically elaborated by a terrestrial crustacean

Orchestia cavimana is a crustacean that cyclically replaces its calcified cuticle during molting cycles in order to grow. Its terrestrial way of life requires storage of calcium during each premolt period, as calcareous concretions, in tubular diverticula of the midgut. During the postmolt period the stored calcium is reabsorbed and is translocated through the storage organ epithelium as calcified small spherules. In a previous study, we sequenced and characterized a remarkable component of the organic matrix of the premolt storage structures, Orchestin, which is a calcium-binding phosphoprotein. In this paper, we analyzed the spatiotemporal expression of the orchestin gene by Northern blotting and in situ hybridization, and its translated product by immunocytochemistry. We found evidence that the gene and the protein are expressed specifically during premolt in the storage organs. More interestingly, we demonstrated that the protein is synthesized also during the postmolt period, as a component of the organic matrix of the calcium resorption spherules. Thus, Orchestin is a matrix component that is synthesized by the same cells to contribute alternately to the elaboration of two different calcifications. These results, in addition to the physical and chemical features of the protein, suggest that Orchestin is probably a key molecule in the calcium carbonate precipitation process leading to the cyclic elaboration of two transitory calcified mineralizations by the crustacean Orchestia.     

Effects of variations in carbonate chemistry on the calcification rates of Madracis auretenra (= Madracis mirabilis sensu Wells, 1973): bicarbonate concentrations best predict calcification rates

Physiological data and models of coral calcification indicate that corals utilize a combination of seawater bicarbonate and (mainly) respiratory CO₂ for calcification, not seawater carbonate. However, a number of investigators are attributing observed negative effects of experimental seawater acidification by CO₂ or hydrochloric acid additions to a reduction in seawater carbonate ion concentration and thus aragonite saturation state. Thus, there is a discrepancy between the physiological and geochemical views of coral biomineralization. Furthermore, not all calcifying organisms respond negatively to decreased pH or saturation state. Together, these discrepancies suggest that other physiological mechanisms, such as a direct effect of reduced pH on calcium or bicarbonate ion transport and/or variable ability to regulate internal pH, are responsible for the variability in reported experimental effects of acidification on calcification. To distinguish the effects of pH, carbonate concentration and bicarbonate concentration on coral calcification, incubations were performed with the coral Madracis auretenra (= Madracis mirabilis sensu Wells, 1973) in modified seawater chemistries. Carbonate parameters were manipulated to isolate the effects of each parameter more effectively than in previous studies, with a total of six different chemistries. Among treatment differences were highly significant. The corals responded strongly to variation in bicarbonate concentration, but not consistently to carbonate concentration, aragonite saturation state or pH. Corals calcified at normal or elevated rates under low pH (7.6–7.8) when the seawater bicarbonate concentrations were above 1800 μm . Conversely, corals incubated at normal pH had low calcification rates if the bicarbonate concentration was lowered. These results demonstrate that coral responses to ocean acidification are more diverse than currently thought, and question the reliability of using carbonate concentration or aragonite saturation state as the sole predictor of the effects of ocean acidification on coral calcification.     

Biomineralization by photosynthetic organisms: Evidence of coevolution of the organisms and their environment?

Biomineralization is widespread among photosynthetic organisms in the ocean, in inland waters and on land. The most quantitatively important biogeochemical role of land plants today in biomineralization is silica deposition in vascular plants, especially grasses. Terrestrial plants also increase the rate of weathering, providing the soluble substrates for biomineralization on land and in water bodies, a role that has had global biogeochemical impacts since the Devonian. The dominant photosynthetic biomineralizers in today's ocean are diatoms and radiolarians depositing silica and coccolithophores and foraminifera depositing calcium carbonate. Abiotic precipitation of silica from supersaturated seawater in the Precambrian preceded intracellular silicification dominated by sponges, then radiolarians and finally diatoms, with successive declines in the silicic acid concentration in the surface ocean, resulting in some decreases in the extent of silicification and, probably, increases in the silicic acid affinity of the active influx mechanisms. Calcium and bicarbonate concentrations in the surface ocean have generally been supersaturating with respect to the three common calcium carbonate biominerals through geological time, allowing external calcification as well as calcification in compartments within cells or organisms. The forms of calcium carbonate in biominerals, and presumably the evolution of the organisms that produce them, have been influenced by abiotic variations in calcium and magnesium concentrations in seawater, and calcium carbonate deposition has probably also been influenced by carbon dioxide concentration whose variations are in part biologically determined. Overall, there has been less biological feedback on the availability of substrates for calcification than is the case for silicification.     

Calcium Dynamics in Land Gastropods

In land gastropods, calcium is precipitated in the shell, inconnective calcium cells which are largely distributed throughthe whole connective tissue, in epithelial calcium cells ofthe digestive gland, and in the calcium gland cells of the skinand the mantle collar. Calcium is taken up from the externalmedium by food and by absorption through the sole skin. To adaptto terrestrial life, these animals have to eliminate appreciableamounts of calcium for their protection and their reproduction.During the egg laying period, a calcium flux occurs throughthe epithelium of the reproductive tract in order to supplythe egg shell and the egg fluids. This egg calcium is takenup by the embryo. The maintenance of a positive calcium balancebetween its uptake and the loss is due to an important reservoirof easily mobilizable calcium in the form of calcium carbonate.This reservoir consists of the connective calcium cells whichare constantly able to accumulate or release calcium as longas calcium is locally available or required. The epithelialcalcium cells of the digestive gland are loaded with calciumphosphate; they are not a major calcium storage compartment,but have an essential function in detoxification. All of thecalcium movement occurring across cell membranes and throughepithelia concerns only calcium ions. All calcium movement canbe regarded either as on-off systems or as reversible systems,both of which are certainly controlled by complex processes